Isolation of microRNA targets by miRNP immunopurification
Abstract
microRNAs (miRNAs) serve as post-transcriptional regulators of gene expression, by guiding effector complexes (miRNPs) to target RNAs. Although considerable progress has been made in computational methods to identify miRNA targets, only a relatively limited assessment of their ability to function in vivo has been reported. Here we describe an alternative approach to miRNA target identification based on a biochemical method for purifying miRNP complexes with associated miRNAs and bound mRNA targets. Microarray analysis revealed a high degree of enrichment for miRNA complementary sites in the 3′UTRs of the miRNP-associated mRNAs. mRNAs specifically associated with an individual miRNA were identified by comparing the miRNP-associated mRNAs from wild-type flies and mutant flies lacking miR-1, and their regulation by the miRNA was validated. This approach provides a means to identify functional miRNA targets based on their physical interaction in vivo.
Keywords
Footnotes
-
Reprint requests to: Stephen M. Cohen, European Molecular Biology Laboratory, Meyerhofstr 1, 69117 Heidelberg, Germany; e-mail: cohen{at}embl.de; fax: 49-6221-387-166.
-
Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.563707.
-
- Received March 15, 2007.
- Accepted May 10, 2007.
- Copyright © 2007 RNA Society